Christian Harwanegg

Microarrayed allergen molecules: diagnostic gatekeepers for allergy treatment

Type I allergy is an immunoglobulin E (IgE)‐mediated hypersensitivity disease affecting more than 25% of the population. Currently, diagnosis of allergy is performed by provocation testing and IgE serology using allergen extracts. This process defines allergen‐containing sources but cannot identify the disease‐eliciting allergenic molecules. We have applied microarray technology to develop a miniaturized allergy test containing 94 purified allergen molecules that represent the most common allergen sources. The allergen microarray allows the determination and monitoring of allergic patients’ IgE reactivity profiles to large numbers of disease‐causing allergens by using single measurements and minute amounts of serum. This method may change established practice in allergy diagnosis, prevention, and therapy. In addition, microarrayed antigens may be applied to the diagnosis of autoimmune and infectious diseases.

Protein microarrays for the diagnosis of allergic diseases: state-of-the-art and future development

Abstract In the emerging field of Functional Proteomics, protein microarrays are considered to be one of the most promising tools for the simultaneous analysis of the a) abundance, b) function, and c) interaction of proteins on a system-wide scale. Resting on the technological grounds of widely used DNA biochips, the great power of microarray-based miniature solid-phase immunoassays lies in their potential to investigate in parallel large numbers of analyte pairs in a variety of biological samples. Consequently, this has fuelled aspirations that protein microarrays may serve as tools for the high-throughput functional investigation of complete proteomes and, moreover, that they will develop into promising candidates for innovative in-vitro diagnostic (IVD) applications. To date, published examples of protein microarrays for IVD purposes have included tests for allergy, autoimmune and infectious diseases. Here, we discuss recent advancements in the development of protein microarrays for the profiling of IgE antibodies in the diagnosis of Type 1-related allergic diseases.

Protein microarrays in diagnosing IgE-mediated diseases: spotting allergy at the molecular level.

Over the last few decades, the prevalence of allergic diseases has increased dramatically in developed nations. The resulting worldwide burden on healthcare systems has provoked a whole series of research initiatives among allergy experts and commercial companies that aim to develop novel tests to improve the diagnostic risk assessment and early preventive treatment of disease. The advent of protein microarray technology has fuelled aspirations of multianalyte immunological applications that permit the simultaneous analysis of huge numbers of disease-related parameters that will hopefully become amenable in the near future. Allergen microarrays have been developed for the monitoring of patient-specific antibody profiles to a previously unknown variety of allergens in a single analytical step. This review describes significant discoveries and developments in allergy research against a background of the increasing prevalence of disease and hence the emerging challenges for national healthcare systems. The development of novel protein microarray-based allergy diagnostic tests is portrayed in concert with the recent advances and benefits of this technology, along with the challenges that must be met by manufacturers in order to succeed with innovative allergen microarrays in a highly competitive market.

Allergen Microarrays for the Diagnosis of Specific IgE Against Components of Cow Milk and Hen Egg in a Multiplex Biochip-Based Immunoassay

Over the last few decades, the prevalence of allergic diseases has increased dramatically in developed nations. The resulting burden on health care systems worldwide has provoked a whole series of research initiatives among allergy experts and commercial companies that aim to develop novel tests to improve the diagnostic risk assessment and early preventive treatment of the disease. The advent of protein microarray technology has inspired the development of miniaturized immunological applications that permit the simultaneous analysis of huge numbers of disease-related parameters. Allergen microarrays have been developed for the monitoring of patient-specific antibody profiles to a previously unknown variety of allergens in a single analytical step. This has been accomplished by the successful adaptation of solid-phase antibody assays for the detection of surface-bound allergens to the microarray format, the development of appropriate assay conditions, and the improvement of software-guided microarray image analysis. Here we report a protocol for the development and analysis of food allergen microarrays.

Protein microarrays for the diagnosis of allergic diseases: State-of-the-art and future development Protein-Biochips fur die Diagnose allergischer Erkrankungen - Stand der Technik und zukunftige Entwicklungen

Abstract In the emerging field of Functional Proteomics, protein microarrays are considered to be one of the most promising tools for the simultaneous analysis of the a) abundance, b) function, and c) interaction of proteins on a system-wide scale. Resting on the technological grounds of widely used DNA biochips, the great power of microarray-based miniature solid-phase immunoassays lies in their potential to investigate in parallel large numbers of analyte pairs in a variety of biological samples. Consequently, this has fuelled aspirations that protein micro-arrays may serve as tools for the high-throughput functional investigation of complete proteomes and, moreover, that they will develop into promising candidates for innovative in-vitro diagnostic (IVD) applications. To date, published examples of protein microarrays for IVD purposes have included tests for allergy, autoimmune and infectious diseases. Here, we discuss recent advancements in the development of protein microarrays for the profiling of IgE antibodies in the diagnosis of Type 1-related allergic diseases. Im ständig wachsenden Forschungsgebiet der funktionellen Proteomik zählen Protein-Biochips aus einer Reihe innovativer Methoden für die quantitative und funktionelle Bestimmung von Proteininteraktionen auf systemischer Ebene heute zu den aussichtsreichsten Kandidaten. Miniaturisierte Festphasen-Immunoassays, die analog zur etablierten DNA-Chip-Technologie entwickelt wurden, entfalten ihr Potential in der hoch parallelen Untersuchung unterschiedlichster Liganden-Paare in biologischem Ausgangsmaterial. Es besteht die berechtigte Hoffnung, dass Protein-Biochips in Zukunft durch den hohen Probendurchsatz und die enorme Zahl der generierten Daten den Bereich der Proteomik revolutionieren werden. Des Weiteren sind sie viel versprechende Kandidaten für neue und innovative in-vitro-Diagnostika (IVD). In mehreren aktuellen Publikationen wurde veranschaulicht, dass Protein-Biochips für die Diagnose von allergischen, autoimmunen oder infektiösen Erkrankungen wertvolle Beiträge leisten können. In diesem Artikel beleuchten wir neue Ansätze und Entwicklungen im Bereich der Diagnostik von IgE-vermittelten Typ 1-Allergien mit Hilfe von Allergen-Biochips.

PreDicta chip-based high resolution diagnosis of rhinovirus-induced wheeze

Rhinovirus (RV) infections are major triggers of acute exacerbations of severe respiratory diseases such as pre-school wheeze, asthma and chronic obstructive pulmonary disease (COPD). The occurrence of numerous RV types is a major challenge for the identification of the culprit virus types and for the improvement of virus type-specific treatment strategies. Here, we develop a chip containing 130 different micro-arrayed RV proteins and peptides and demonstrate in a cohort of 120 pre-school children, most of whom had been hospitalized due to acute wheeze, that it is possible to determine the culprit RV species with a minute blood sample by serology. Importantly, we identify RV-A and RV-C species as giving rise to most severe respiratory symptoms. Thus, we have generated a chip for the serological identification of RV-induced respiratory illness which should be useful for the rational development of preventive and therapeutic strategies targeting the most important RV types.Rhinovirus (RV) infections can trigger acute exacerbations of respiratory diseases. Here, Niespodziana et al. develop a PreDicta chip that identifies the culprit RV strain from small blood samples and show that RV-A and RV-C strains are associated with most severe symptoms.